Wadsworth-Ktl Anaerobic Bacteriology Manual

by Jousimies-Somer, Hannele R.; Summanen, Paula; Ditron, Diane M.; Baron, Ellen Jo; Wexler, Hannah M.; Finegold, Sydney M.

• ISBN13: 9780898632095
• ISBN10: 0898632099
• Edition: 6th
• Format: Paperback
• Copyright: 2002-06-01
• Publisher: Start Now Pr

Summary

Anaerobic bacteria may be involved in virtually any type of bacterial infection at any site in the body. Often they are part of a mixed flora, but some infections involve only anaerobes. Despite frequently being the principal cause of infection, anaerobes may be readily overlooked in infectious processes. Ironically, this is partly because of the antimicrobials available with excellent activity against anaerobes, and partly because of the failure to identify anaerobes -- the latter sometimes is a result of reduced budgets in clinical laboratories. Unfortunately, these antimicrobials can have serious consequences, including increased expense -- many of these antibiotics are expensive -- and the real risk of increased resistance to these agents. Of course, the most important immediate effect may be harm to the patient. There is a critical need for the Wadsworth-KTL Anaerobic Bacteriology Manual. Although the authors emphasize practical approaches to anaerobic bacteriology for clinical laboratories, they provide additional information on more specialized techniques and procedures for the study and identification of more fastidious organisms. In this sixth edition, you'll find new identification methods, with new color-coded flow charts that illustrate procedures succinctly. Furthermore, the authors provide collection and transport techniques, and susceptibility data that are essential to clinicians and laboratory personnel. New, detailed information on recent taxonomic changes of anaerobic bacteria is also provided. Book jacket.

Table of Contents

Preface	xiii
Introduction to Anaerobic Bacteriology	1	(22)
The Indigenous Flora	2	(4)
The Oral Flora	3	(1)
The Gastrointestinal Flora	3	(3)
The Flora of the Female Genital Tract	6	(1)
Optimal Anaerobic Bacteriology in the Face of Restricted Resources	6	(2)
Clinical Background	8	(15)
Incidence of Anaerobes in Infection	8	(1)
Clues to Anaerobic Infection	9	(1)
Taxonomic Changes	9	(14)
Specimen Collection and Anaerobic Culture Techniques	23	(20)
Specimen Collection	23	(8)
Principle	23	(1)
Specimen Collection Methods	24	(7)
Specimen Transport	31	(3)
Anaerobic Culture Techniques	34	(9)
Blood Culture Techniques	34	(2)
Anaerobic Incubation Methods	36	(6)
Common Pitfalls in Specimen Collection and Culture Techniques	42	(1)
Processing Clinical Specimens and Isolation Procedures	43	(12)
Initial Processing Procedure	43	(6)
Direct Examination	43	(3)
Specimen Preparationand Inoculation	46	(2)
Incubation of Cultures	48	(1)
Examination of Cultures and Isolation of Anaerobes	49	(6)
Examination of Primary Plates	49	(1)
Subculture of Isolates	50	(4)
Common Pitfalls in Specimen Processing and Isolation Procedures	54	(1)
Preliminary Identification Methods	55	(20)
General Considerations	55	(2)
Presumptive Identification	57	(18)
Preliminary Examination of All Isolates	57	(8)
Screening Tests for Selected Isolates	65	(1)
Gram-Negative Organisms	66	(4)
Gram-Positive Organisms	70	(3)
Commonly Encountered Errors in Identification	73	(2)
Rapid, Cost-Efficient Identification Methods Using Preformed Enzyme Tests	75	(6)
General Considerations	75	(1)
Identification Using Preformed Enzyme Tests	75	(6)
Common Pitfalls Associated with Preformed Enzyme Tests	79	(2)
Advanced Identification Methods	81	(52)
General Considerations	81	(1)
Advanced Identification	81	(52)
Gram-Negative Bacilli	81	(3)
Bacteroides fragilis Group	84	(2)
Nonpigmented Prevotella and Other Gram-Negative Rods	86	(6)
Pigmented Gram-Negative Rods	92	(6)
Fusobacteria	98	(5)
Gram-Negative Cocci	103	(1)
Gram-Positive Cocci	103	(3)
Gram-Positive Bacilli	106	(8)
Gram-Positive Nonsporeforming Bacilli	114	(16)
Identification Systems	130	(3)
Laboratory Tests for Diagnosis of Clostridium Difficile Enteric Disease	133	(10)
General Priniciples	133	(2)
Laboratory Procedures	135	(8)
Specimen Collection for All Procedures	135	(1)
Culture Procedures	135	(3)
Cytotoxin Testing Using Commercial Systems	138	(2)
Enzyme Immunoassay Tests for Enterotoxin and/or Cytotoxin and/or Cellular Antigen	140	(3)
Susceptibility Testing of Anaerobic Bacteria	143	(22)
Resistance Mechanisms in Anaerobic Bacteria	143	(1)
Overview of Current Susceptibility Patterns	143	(2)
Summary of Data Collected at the Wadsworth Anaerobe Laboratory	143	(1)
Summary of Data From Other Laboratories	144	(1)
General Considerations	145	(4)
Procedures for Testing Susceptibility	149	(16)
Indications for Susceptibility Testing	149	(3)
Agents to be Tested	152	(1)
Preparation of Inoculum	152	(1)
Preparation of Antimicrobial Stock Solutions and Dilutions	153	(1)
Addition of Growth Supplements to Antimicrobial Susceptibility Test Media	153	(1)
Quality Control Standards	154	(1)
Reference Agar Dilution Test Procedure	155	(5)
Broth Microdilution Test	160	(2)
MBC Determination	162	(1)
Broth Macrodilution Tests	162	(1)
Other Methods of Susceptibility Testing	162	(1)
Etest	162	(1)
Spiral Gradient Endpoint System	163	(1)
β-Lactamase Testing	164	(1)
APPENDIX A SPECIAL PROCEDURES FOR SPECIMEN COLLECTION AND RESEARCH STUDIES	165	(22)
Laboratory Handling of Oral Site Specimens to Avoid Normal Oral Flora	165	(4)
Principle	165	(1)
Saliva Collection	166	(1)
Supragingival Plaque Specimen Collection	166	(1)
Subgingival Plaque Specimen Collection	167	(1)
Periimplant Plaque Specimen Collection	167	(1)
Processing Oral Specimens in the Laboratory	168	(1)
Anaerobes in Bite Wound Infections	169	(1)
Respiratory Tract Specimens Suitable for Anaerobic Culture	170	(3)
Principle	170	(2)
Processing the PBC Specimen	172	(1)
Laboratory Processing of the PBC or BAL Specimen	172	(1)
Collection of Endometrial Specimens Using the Pipelle Protected Suction Curette	173	(1)
Principle	173	(1)
Specimen Collection	174	(1)
Specimen Processing	174	(1)
Vaginal Flora	174	(4)
Vaginal Wash	175	(1)
Calibrated Loop	175	(1)
Processing Vaginal Specimens	175	(2)
Bacterial Vaginosis	177	(1)
Quantitative Studies of Intestinal Flora: Collection and Transport	178	(4)
Fecal Specimens	178	(1)
Other Specimens of Intestinal Contents	178	(1)
Processing of Specimens	179	(3)
Identification of Isolates	182	(1)
Descriptions of Typical Colonies on Selective Media for Anaerobes	182	(5)
Bacteroides Bile Esculin Agar (BBE)	183	(1)
Blood and Liver (BL) and Bifidobacterium- Selective (BS) Agars	183	(1)
Cadmium Sulfate Flouride Acriflavine Tellurite Agar (CFAT)	183	(1)
Cefoxitin Cycloserine Egg Yolk (CCEY)	183	(1)
Cycloserine Cefotoxin Fructose Agar (CCFA)	184	(1)
Egg Yolk Agar (EYA)	184	(1)
Fusobacterium-Selective Medium (JVN) and Fusobacterium neomycin vancomycin (NV) Agar	184	(1)
Kanamycin Vancomycin Laked Blood (KVLB) Agar	184	(1)
Lactobacillus-Selective Medium (LBS)	185	(1)
MRS (de Man, Rogosa, Sharpe) Medium	185	(1)
Rifampin (RIF) Blood Agar	185	(1)
Veillonella Neomycin Agar	185	(1)
Trypticase Soy Bacitracin Vancomycin (TSBV) Agar	185	(2)
APPENDIX B GAS-LIQUID CHROMATOGRAPHY FOR METABOLIC ENDPRODUCT AND CELLULAR FATTY ACID METHYL ESTER ANALYSIS	187	(4)
Short-Chain Fatty Acid Analysis	187	(2)
Chromatographs, Columns, and Operating Conditions	187	(1)
Preparation of Samples for Analysis	188	(1)
Volatile Fatty Acids	188	(1)
Nonvolatile Fatty Acids	188	(1)
Methyl Ester (Fame) Analysis	189	(2)
Procedure	189	(2)
APPENDIX C BIOCHEMICAL TEST PROCEDURES	191	(16)
Bile	191	(1)
Carbohydrate Fermentation	192	(1)
Catalase Test	192	(1)
Catalyst Regeneration	193	(1)
Conversion of Lactate to Propionate	193	(1)
Conversion of Threonine to Propionate	193	(1)
Desulfoviridin Test	194	(1)
Esculin Hydrolysis	194	(1)
Flagella Stain	195	(1)
Fluorescence	195	(1)
Gelatin Liquefaction	196	(1)
Growth Stimulation Test	196	(1)
Indole Production (Tryptophanase)	197	(1)
Lecithinase	198	(1)
Lipase	199	(1)
Milk	199	(1)
Motility	200	(1)
Nitrate Reduction Test	200	(1)
Pras Biochemical Inoculation	200	(1)
Rapid Enzyme Tests	201	(1)
Reverse-CAMP Test	202	(1)
Special-Potency Disk Identification	203	(1)
Spore Test	203	(1)
Trypsinlike Activity	204	(1)
Urease	204	(3)
APPENDIX D PREPARATION OF MEDIA AND REAGENTS	207	(28)
Quality Control Procedures	207	(2)
Media and Reagents	209	(26)
Arginine (10%)	209	(1)
Bacteroides Bile Esculin Agar (BBE)	210	(1)
BL (Blood and Liver) Agar	210	(1)
BS (Bifidobacterium-Selective) Agar	211	(1)
Brucella Blood Agar	211	(1)
Brucella Blood Agar for Susceptibility Testing	212	(1)
Brucella Broth	212	(1)
Cadmium Flouride Acriflavine Tellurite Agar (CFAT)	213	(1)
Cefotoxin Cycloserine Egg Yolk Agar (CCEY)	213	(1)
Chocolate Bacitracin Agar	214	(1)
Cycloserine Cefotoxin Fructose Agar (CCFA)	214	(1)
Egg Yolk Agar (EYA)	215	(1)
Egg Yolk Emulsion	216	(1)
Ehrlich's Reagent	216	(1)
Ferric Ammonium Citrate (1%)	217	(1)
Fildes Enrichment	217	(1)
Flagella Stain (Ryu)	217	(1)
Formate Fumarate Additive	217	(1)
Fusobacterium-Selective (JVN) Agar	218	(1)
Fusobacterium Neomycin Vancomycin (NV) Agar	218	(1)
Hemin Solution (5 mg/ml)	219	(1)
Kanamycin Soluton (100 mg/ml)	219	(1)
Kanamycin Vancomycin Laked Blood Agar (KVLB)	220	(1)
Lactobacillus-Selective Medium (LBS)	220	(1)
Laked Blood	220	(1)
Magnesium Sulfate (10%)	221	(1)
McFarland Standards	221	(1)
0.5 McFarland Standard	222	(1)
Methanobacterium Medium (Pras Roll Tubes)	222	(1)
Man-Rogosa-Sharpe Medium (MRS)	223	(1)
Neomycin Solution (100 mg/ml)	224	(1)
Nessler Reagent	224	(1)
Nitrate Disks	224	(1)
Nitrate Reagents	225	(1)
Oxgall (40%)	225	(1)
Paradimethylaminocinnamaldehyde	225	(1)
Phenylethyl Alcohol Blood Agar (PEA)	226	(1)
Pyruvate (20%)	226	(1)
Resazurin	226	(1)
Rifampin (1 mg/ml)	227	(1)
Rifampin Blood Agar	227	(1)
Ringer's Solution	227	(1)
One-Quarter Strength Ringer's Solution with Metaphosphate	228	(1)
Ringer's Dilution Solution	228	(1)
Skim Milk (20%)	228	(1)
Sodium Bicarbonate (20 mg/ml)	229	(1)
Sodium Polyanethol Sulfonate (SPS) Disks	229	(1)
TE Buffer (50 mM)	229	(1)
Thioglycolate, Supplemented	230	(1)
Tryptic Soy Serum Bacitracin Vancomycin (TSBV) Agar	230	(1)
Tween-80	231	(1)
Urea Broth	231	(1)
Vancomycin Stock Solution (7.5 mg/ml)	232	(1)
Veillonella Neomycin Agar (VNA)	232	(1)
Vitamin K1 Solution (1 mg/ml)	232	(1)
VPI Salts Solution	233	(2)
APPENDIX E STOCKING AND SHIPPING CULTURES	235	(2)
Stocking Cultures	235	(1)
Shipping Cultures	235	(2)
APPENDIX F SOURCES OF SUPPLIES	237	(4)
Media and Reagents	237	(2)
Equipment and Supplies	239	(1)
Reference Cultures	240	(1)
References	241	(26)
Index	267